Identification of the endophilins (SH3p4/p8/p13) as novel binding partners for the beta1-adrenergic receptor.

Several G-protein coupled receptors, such as the beta1-adrenergic receptor (beta1-AR), contain polyproline motifs within their intracellular domains. Such motifs in other proteins are known to mediate protein-protein interactions such as with Src homology (SH)3 domains. Accordingly, we used the proline-rich third intracellular loop of the beta1-AR either as a glutathione S-transferase fusion protein in biochemical "pull-down" assays or as bait in the yeast two-hybrid system to search for interacting proteins. Both approaches identified SH3p4/p8/p13 (also referred to as endophilin 1/2/3), a SH3 domain-containing protein family, as binding partners for the beta1-AR. In vitro and in human embryonic kidney (HEK) 293 cells, SH3p4 specifically binds to the third intracellular loop of the beta1-AR but not to that of the beta2-AR. Moreover, this interaction is mediated by the C-terminal SH3 domain of SH3p4. Functionally, overexpression of SH3p4 promotes agonist-induced internalization and modestly decreases the Gs coupling efficacy of beta1-ARs in HEK293 cells while having no effect on beta2-ARs. Thus, our studies demonstrate a role of the SH3p4/p8/p13 protein family in beta1-AR signaling and suggest that interaction between proline-rich motifs and SH3-containing proteins may represent a previously underappreciated aspect of G-protein coupled receptor signaling.

Regions of the alpha 1-adrenergic receptor involved in coupling to phosphatidylinositol hydrolysis and enhanced sensitivity of biological function.

Regions of the hamster alpha 1-adrenergic receptor (alpha 1 AR) that are important in GTP-binding protein (G protein)-mediated activation of phospholipase C were determined by studying the biological functions of mutant receptors constructed by recombinant DNA techniques. A chimeric receptor consisting of the beta 2-adrenergic receptor (beta 2AR) into which the putative third cytoplasmic loop of the alpha 1AR had been placed activated phosphatidylinositol metabolism as effectively as the native alpha 1AR, as did a truncated alpha 1AR lacking the last 47 residues in its cytoplasmic tail. Substitutions of beta 2AR amino acid sequence in the intermediate portions of the third cytoplasmic loop of the alpha 1AR or at the N-terminal portion of the cytoplasmic tail caused marked decreases in receptor coupling to phospholipase C. Conservative substitutions of two residues in the C terminus of the third cytoplasmic loop (Ala293----Leu, Lys290----His) increased the potency of agonists for stimulating phosphatidylinositol metabolism by up to 2 orders of magnitude. These data indicate (i) that the regions of the alpha 1AR that determine coupling to phosphatidylinositol metabolism are similar to those previously shown to be involved in coupling of beta 2AR to adenylate cyclase stimulation and (ii) that point mutations of a G-protein-coupled receptor can cause remarkable increases in sensitivity of biological response.

Report on Archaeological Investigations Conducted at the St. Mary's Site (18AP45), 107 Duke of Gloucester Street, Annapolis, Maryland, 1987-1990

In celebration of the 250th anniversary of the birth of Charles Carroll of Carrollton, Archaeology in Annapolis was invited to excavate the Carroll House and garden on 107 Duke of Gloucester Street in Annapolis, Maryland. The site, named the St. Mary's Site (18AP45) for the Catholic church on the property, is currently owned by the Redemptorists, a Roman Catholic congregation of priests and brothers who have occupied the site since 1852. Prior to the Redemptorists' tenure, the site was owned by the Carroll family from 1701-1852 and is perhaps best known as the home of Charles Carroll of Carrollton (1737-1832), signer of the Declaration of Independence. Excavations at the site were conducted during four consecutive summer seasons from 1987-1990. The investigation focused on three research questions. The first line of inquiry were questions surrounding the dating, architectural configuration, and artifact deposits of the "frame house," a structure adjoining the west wall of the brick Carroll House via a "passage" and later a three story addition. The frame house was partially demolished in the mid-nineteenth century but the construction was thought to pre-date the brick portion of the house. The second research question was spurred by documentary research which indicated that the property might have been the location of Proctor's Tavern, a late 17th-century tavern which served as the meeting place of the Maryland Provincial Assembly. Archaeological testing hoped to determine its location and, if found, investigate Annapolis' early Euro-American occupation. The third research question focused on the landscape of the site as it was shaped by its occupants over the past three hundred years. The research questions included investigating the stratigraphy, geometry, and architectural and planting features of Charles Carroll of Carrollton's terraced garden built during the 1770s, and investigating the changes to the landscape made by the Redemptorists in the nineteenth and twentieth centuries. While no structural evidence associated with Proctor’s Tavern was uncovered during limited excavations along Spa Creek, the historic shore of Spa Creek was identified, buried beneath deep fill deposits laid down during construction of the Carroll Garden. Features and deposits associated with this period likely remain intact in a waterlogged environment along the southeastern sea wall at the St. Mary’s Site. Evidence of extensive earth moving by Carroll is present in the garden and was identified during excavation and coring. This strongly suggests that the garden landscape visible at the St. Mary’s Site is the intact Carroll Garden, which survives beneath contemporary and late nineteenth century strata. The extant surviving garden should be considered highly sensitive to ground-disturbing activities, and is also highly significant considering demonstrable associations with the Carroll family. Other garden-related features were also discovered, including planting holes, and a brick pavilion or parapet located along Spa Creek to the south of the site. The Duke of Gloucester Street wall was shown to be associated with the Carroll occupation of the site. Finally, intensive archaeological research was directed at the vicinity of a frame house constructed and occupied by the Carrolls to the east of the existing brick house, which was replaced by the Redemptorists in the nineteenth century with a greenhouse. These superimposed buildings were documented in detail and remain highly significant features at the St. Mary’s Site.

"It is Quietly Chaotic. It Confuses Time"*: Final Report of Excavations at the Bordley-Randall Site in Annapolis, Maryland, 1993-1995 (18AP50)

*This extract is from Gay P. Crowther's description of the Randall Court pathway (Cowther 1985).

The Oud Across Arabic Culture

This study is a compilation and compendium of information on the oud, the most important instrument in Arabic classical music. It has grown out of my own long-time involvement in studying and playing the oud, and in particular out of my interest in the lack of sources and knowledge available to the vast majority of oud players and researchers, as well as for the readers. My own path started from an intensive study of the oud, which included exposure to several treaties; some housed in museums around the globe, and some only available in the Arabic language. The study combines archival research (including Arabic poetry and pre-Islamic Era and medieval treaties), symbolism, new archaeological discoveries, field interviews, and analysis of existing scholarship, and draws on my professional performance experience for detailed stylistic analysis of the oud's performance practice and its historical development. The study consists of participant observation, personal performance, and interviews conducted in person, via telephone, and/or via e-mail, according to the choice of the performers. The performers have been selected from networks of musicians who perform regularly at lounges, concert halls, and private events. These performers have been chosen according to their musical knowledge, technical skill, experience, and activity in Arabic music and oud performance. Chapter one deals with the purpose of this study and the methods of investigation, as well as giving a brief overview of the history of the oud. In addition, there will be an introduction to the Arabic musical system (mâqâm), which is primarily based on the mechanics and sound production of the oud. Chapter two deals with the oud in Arabic sources: the first source is Arabic poetry in the pre-Islamic Era. The second source is Arabic poetry in the medieval era, in which I found a significant number of poets who allude to the oud, providing accurate descriptions of the player, singers, and the scenes within the contexts of oud performance. The third source is the Arab scholars' intensive treatises with meticulous accounts of the instrument's apparatii, including descriptions and measurements of the parts, strings, and tuning. While chapter three deals with the classification, the development of the oud, chapter four deals with topics such as: the symbolism of the oud and its relation to cosmology, astronomy, mathematics and anatomy. In most of the pertinent Arabic writings, philosophers mention a significant correlation between the oud and the other sciences. Chapter five deals with recreating the performance practice of the oud. A case study of the oud performers focuses on their style, technique, training, and personal experiences. Topics such as improvisation and ornamentation, the oud in the Arabic musical ensemble, the social uses and functions, and gender in musical performance practices will be included in detailed analysis. Other important topics will be analyzed such as traditional vs. modern technique, and the repertoire of the oud. Specifically, in regard to technique, the study outline the style of the music, the role of the oud in Arabic ensembles, the function of the oud in music composition, and the form of the ensembles in Arabic performance and practice.

Memory and coping with stress: the relationship between cognitive-emotional distinctiveness, memory valence, and distress.

Cognitive-emotional distinctiveness (CED), the extent to which an individual separates emotions from an event in the cognitive representation of the event, was explored in four studies. CED was measured using a modified multidimensional scaling procedure. The first study found that lower levels of CED in memories of the September 11 terrorist attacks predicted greater frequency of intrusive thoughts about the attacks. The second study revealed that CED levels are higher in negative events, in comparison to positive events and that low CED levels in emotionally intense negative events are associated with a pattern of greater event-related distress. The third study replicated the findings from the previous study when examining CED levels in participants' memories of the 2004 Presidential election. The fourth study revealed that low CED in emotionally intense negative events is associated with worse mental health. We argue that CED is an adaptive and healthy coping feature of stressful memories.

Exploring Musical Diversity in the Collaborative Repertoire from 1880 through 1963

It is essential in musical performance not only to convey the unique language of the composers but also to approach each composition from the perspective of its style. During the 20th century, diverse musical idioms co-existed, sometimes mixing or fusing, yet retaining recognizable characteristics and thereby remaining distinctive. This dissertation explores myriad examples from Late Romanticism/Post- Romanticism, Naturalism, Neo-Classicism, Nationalism and Impressionism composed during this unusually rich period. In order to explore a broad range of collaborative repertoire and to deepen my knowledge of the styles and performance practices relating to these pieces, I studied and performed the repertoire with pianist Eunae Baik–Kim, clarinetist Jihoon Chang, and singers Joshua Brown and Young Joo Lee. The first program featured Post-Romantic, Neo-Classic and Impressionist two-piano works composed by Debussy, Rachmaninoff, and Stravinsky. Each of the three composers used their own distinctive harmonies, rhythms, melodic inventions, pedaling and figurations. In all of the works, both piano parts were densely interwoven, having equal importance. Lied and operatic aria was the focus of the second recital. Brahms’ Vier Ernste Gesänge Op. 121, Ravel’s Don Quichotte a Dulcineé and Italian, French and German operatic arias were the examples of Post-Romanticism and Nationalism. The representative composers were Verdi, Massenet, Korngold, Leoncavallo, Ravel and Wagner. Despite the fact that all of the repertoire was written in traditional musical forms, the composers’ unique voices mark each work as belonging to a particular genre. The third recital focused on Post-Romantic and Impressionistic music written for clarinet and piano: the Première Rhapsodie by Debussy, the Sonata by Poulenc and Brahms’ Sonata in F minor Op. 120, No. 1. These works, although profoundly different in style, share elements of simplicity, clarity and elegance as well as technical virtuosity, articulation and profound musical depth. The three recitals which comprise this dissertation project were performed at the University of Maryland Gildenhorn Recital Hall on February 27, 2010, October 25, 2010, and January 31, 2011. The recitals were recorded on compact disc and are archived within the Digital Repository at the University of Maryland (DRUM).

JOACHIM, BRAHMS, AND SCHUMANN: THE FLOURSHING OF NINETEENTHCENTURY VIOLIN REPERTOIRE

The cultivation of violin repertoire and its ultimate dominance of the late Nineteenth-Century orchestral library are best examined through the analysis and study of the works of Joseph Joachim (1831-1907), Johannes Brahms (1833-1897), and Robert Schumann (1810-1856). Each of these men, in their own right, made significant contributions to the development of violin repertoire during the latter half of the Nineteenth Century. Yet their achievements were also the result of a collaborative effort and shared influence, the absence of which would have yielded a diminutive musical landscape, especially in the concerti of Brahms and Joachim. This dissertation explores Joachim’s technical dexterity and its influence on Brahms and Schumann, and further studies the vital role Brahms and Schumann played in forming Joachim as both composer and editor. The pieces examined in this dissertation evidence the significant influence each of these composers shared. Three chamber compositions stand as guideposts in the analysis and establish a stylistic foundation to collaborative efforts among Joachim, Brahms, and Schumann. The preliminary recital focuses on these chamber pieces which illustrate the individual style of each composer, featuring Joachim’s Romance, Op. 2, written between 1848 and 1852, Schumann’s Second Sonata, Op. 121, written in 1851, and Brahms’ Second Sonata, Op. 100, written in 1886. A second performance includes the enigmatic F-A-E Sonata of Brahms, Schumann, and Albert Deitrich, Schumann’s pupil, as well as Joachim’s Second Concerto. A collaborative effort, the F-A-E Sonata represents Brahms’ and Schumann’s efforts to write with Joachim’s unbridled style and technique in mind. An even greater musical offering, Joachim’s Second Concerto, a gift to Brahms in 1860, has been called the “Holy Grail” of concertos, and is considered the preeminent display of Joachim’s creative genius by incorporating demanding technical challenges and Hungarian-like overtones. The third and final program features Brahms’ Concerto in D Major. A fitting response to Joachim’s gift 17 years later, Brahms’ Concerto was written for Joachim at the height of his career, bearing the performer’s stylistic elements throughout. The recital also includes various Hungarian Dances by Brahms. While the Dances were not original to Brahms, they further illustrate the underlying idiom of Hungarian folk music in both Brahms’ and Joachim’s art.

THE ART OF TRANSCRIBING FOR HARPSICHORD

A relatively unexplored area of the harpsichord repertoire is the group of transcriptions made by J.S. Bach (1685-1750), Jean Henry d'Anglebert (1629-1691), and Jean-Baptiste Forqueray (1699-1782). These transcriptions are valuable and worth exploring and performing. Studying them provides unique insights into their composer‘s musical thinking. By comparing transcriptions with their original sources, the transcriber's decisions and priorities can be observed. The performance component of this dissertation comprises three recitals. The first features works of Johann Sebastian Bach: two transcriptions of violin concerti by Antonio Vivaldi (1678-1741), and two transcriptions of trio sonatas by Johann Adam Reinken (1643-1722). The most salient feature of Bach‘s transcriptions is his addition of musical material: ornamenting slow movements, adding diminutions and idiomatic keyboard figurations throughout, and recomposing and expanding fugal movements. The second recital features works of Jean Henry d'Anglebert and Jean-Baptiste Forqueray, two French composer/performers. From d'Anglebert‘s many transcriptions, I assembled two key-related suites: the first comprised of lute pieces by Ennemond Gaultier (c. 1575-1651), and the second comprised of movements from operas by Jean-Baptiste Lully (1632-1687). Forqueray's transcriptions are of suites for viola da gamba and continuo, composed by his father, Antoine Forqueray (1671-1745). Creative and varied ornamentation, along with the style brisé of arpeggiated chords, are the most important features of d‘Anglebert‘s transcriptions. Forqueray‘s transcriptions are highly virtuosic and often feature the tenor and bass range of the harpsichord. The third recital features my own transcriptions: the first suite for solo cello by J.S. Bach, excerpts from the opera La Descente d’Orphée aux Enfers by Marc-Antoine Charpentier (1643-1704), and two violin pieces by Nicola Matteis (fl. c. 1670-c. 1698). In these transcriptions, I demonstrate what I have learned from studying and performing the works in the first two recitals. These recitals were performed in the Leah Smith Hall at the University of Maryland on May 4, 2010; May 11, 2010; and October 7, 2010. They were recorded on compact discs and are archived within the Digital Repository at the University of Maryland (DRUM).

The Evolution of Rachmaninov's Pianistic Style from 1892 to 1941 as Seen in Selected Original Works and Transcriptions for Piano

Unlike some other major composer-pianists (Franz Liszt and Alexander Scriabin being the most notable examples), Sergei Rachmaninov did not experience any dramatic changes in his compositional style; one can, however, notice certain stylistic differences between piano works composed during different periods of his long creative life. This dissertation argues that a clear evolution of Rachmaninov's pianistic style through his three creative periods can be traced by examining a selection of his compositions, from his first significant cycle for piano, 5 Marceaux de fantaisie, Op. 3 (1892), all the way through to his last piano work, a transcription of P.I.Tchakovsky's Lullaby, Op. 16, No. 1 (1941). Rachmaninov's life as a composer can be divided into three periods. His early period was ended abruptly by the disastrous premiere ofhis First Symphony in 1897, which caused a deep psychological crisis in the life of the young composer. Piano works of this period are often characterized by relatively simple homophonic texture, when Rachmaninov was clearly influenced by some of his Russian predecessors, most notably Tchaikovsky. His second and most productive period, also known as the period of"Russian maturity," started in 1900, when he began working on the Second Suite for two pianos, Op. 17, and the Second Concerto, Op. 18; this phase ended with the Russian Bolshevik revolution of 1917. Works of this time exhibit a mature style of piano writing, with rich, virtuosic - often considered excessive by many critics - texture and ever-increasing use of chromatic harmonies. Rachmaninov's works of the third period are limited in number owing to the composer's preoccupation with a career as pianist. Original works for piano now give way to revisions of earlier compositions and transcriptions: Rachmaninov's piano writing becomes more efficient and economical without losing any of its virtuosic brilliance. This dissertation project examines in detail, over the course of three piano recitals, a variety of works composed during the "Russian maturity" period, from several Preludes from Op. 23 (1903), the first major cycle for solo piano of the period, to 9 Etudes-Tableaux, Op.39 (1917), the last one; the early period is represented by 3 of 5 Marceaux defantaisie, Op. 3 (1892), while the late period is shown through several piano transcriptions and revised versions of the remaining 2 pieces from Op. 3.

Human Non-neutralizing HIV-1 Envelope Monoclonal Antibodies Limit the Number of Founder Viruses during SHIV Mucosal Infection in Rhesus Macaques.

HIV-1 mucosal transmission begins with virus or virus-infected cells moving through mucus across mucosal epithelium to infect CD4+ T cells. Although broadly neutralizing antibodies (bnAbs) are the type of HIV-1 antibodies that are most likely protective, they are not induced with current vaccine candidates. In contrast, antibodies that do not neutralize primary HIV-1 strains in the TZM-bl infection assay are readily induced by current vaccine candidates and have also been implicated as secondary correlates of decreased HIV-1 risk in the RV144 vaccine efficacy trial. Here, we have studied the capacity of anti-Env monoclonal antibodies (mAbs) against either the immunodominant region of gp41 (7B2 IgG1), the first constant region of gp120 (A32 IgG1), or the third variable loop (V3) of gp120 (CH22 IgG1) to modulate in vivo rectal mucosal transmission of a high-dose simian-human immunodeficiency virus (SHIV-BaL) in rhesus macaques. 7B2 IgG1 or A32 IgG1, each containing mutations to enhance Fc function, was administered passively to rhesus macaques but afforded no protection against productive clinical infection while the positive control antibody CH22 IgG1 prevented infection in 4 of 6 animals. Enumeration of transmitted/founder (T/F) viruses revealed that passive infusion of each of the three antibodies significantly reduced the number of T/F genomes. Thus, some antibodies that bind HIV-1 Env but fail to neutralize virus in traditional neutralization assays may limit the number of T/F viruses involved in transmission without leading to enhancement of viral infection. For one of these mAbs, gp41 mAb 7B2, we provide the first co-crystal structure in complex with a common cyclical loop motif demonstrated to be critical for infection by other retroviruses.

BEYOND THE LIBRETTO: HOW THE COMPOSER GUIDES THE CREATION OF CHARACTER THREE CASE STUDIES

When performing in opera, a singer portrays a character. A libretto is used as the principal resource for the research. Music can also reveal insights into the composer’s ideas regarding characterization. This performance dissertation examines how musical devices such as genre, texture, meter, melody, instrumentation and form can be used to inform choices of characterization. Three roles from diverse operas were examined and performed. The first role, Estelle Oglethorpe in Later the Same Evening (2007) by John Musto (b 1954) was performed November 15, 16, 17, 18 2007. The second role, Dorabella in Così fan tutte (1789) by Wolfgang Amadeus Mozart (1756-1791) was performed April 20, 25, 27, 2008. The third role, Olga in Eugene Onegin (1878) by Pyotr Ilyich Tchaikovsky (1840-1893) was performed on April 19, 2009. All operas were presented by the University of Maryland Opera Studio at the Ina and Jack Kay Theater in the Clarice Smith Performing Arts Center, University of Maryland College Park. DVD recordings of all performances can be found in the University of Maryland library system.

Micro-Anatomical Quantitative Imaging Towards Enabling Automated Diagnosis of Thick Tissues at the Point of Care

Histopathology is the clinical standard for tissue diagnosis. However, histopathology has several limitations including that it requires tissue processing, which can take 30 minutes or more, and requires a highly trained pathologist to diagnose the tissue. Additionally, the diagnosis is qualitative, and the lack of quantitation leads to possible observer-specific diagnosis. Taken together, it is difficult to diagnose tissue at the point of care using histopathology.

Several clinical situations could benefit from more rapid and automated histological processing, which could reduce the time and the number of steps required between obtaining a fresh tissue specimen and rendering a diagnosis. For example, there is need for rapid detection of residual cancer on the surface of tumor resection specimens during excisional surgeries, which is known as intraoperative tumor margin assessment. Additionally, rapid assessment of biopsy specimens at the point-of-care could enable clinicians to confirm that a suspicious lesion is successfully sampled, thus preventing an unnecessary repeat biopsy procedure. Rapid and low cost histological processing could also be potentially useful in settings lacking the human resources and equipment necessary to perform standard histologic assessment. Lastly, automated interpretation of tissue samples could potentially reduce inter-observer error, particularly in the diagnosis of borderline lesions.

To address these needs, high quality microscopic images of the tissue must be obtained in rapid timeframes, in order for a pathologic assessment to be useful for guiding the intervention. Optical microscopy is a powerful technique to obtain high-resolution images of tissue morphology in real-time at the point of care, without the need for tissue processing. In particular, a number of groups have combined fluorescence microscopy with vital fluorescent stains to visualize micro-anatomical features of thick (i.e. unsectioned or unprocessed) tissue. However, robust methods for segmentation and quantitative analysis of heterogeneous images are essential to enable automated diagnosis. Thus, the goal of this work was to obtain high resolution imaging of tissue morphology through employing fluorescence microscopy and vital fluorescent stains and to develop a quantitative strategy to segment and quantify tissue features in heterogeneous images, such as nuclei and the surrounding stroma, which will enable automated diagnosis of thick tissues.

To achieve these goals, three specific aims were proposed. The first aim was to develop an image processing method that can differentiate nuclei from background tissue heterogeneity and enable automated diagnosis of thick tissue at the point of care. A computational technique called sparse component analysis (SCA) was adapted to isolate features of interest, such as nuclei, from the background. SCA has been used previously in the image processing community for image compression, enhancement, and restoration, but has never been applied to separate distinct tissue types in a heterogeneous image. In combination with a high resolution fluorescence microendoscope (HRME) and a contrast agent acriflavine, the utility of this technique was demonstrated through imaging preclinical sarcoma tumor margins. Acriflavine localizes to the nuclei of cells where it reversibly associates with RNA and DNA. Additionally, acriflavine shows some affinity for collagen and muscle. SCA was adapted to isolate acriflavine positive features or APFs (which correspond to RNA and DNA) from background tissue heterogeneity. The circle transform (CT) was applied to the SCA output to quantify the size and density of overlapping APFs. The sensitivity of the SCA+CT approach to variations in APF size, density and background heterogeneity was demonstrated through simulations. Specifically, SCA+CT achieved the lowest errors for higher contrast ratios and larger APF sizes. When applied to tissue images of excised sarcoma margins, SCA+CT correctly isolated APFs and showed consistently increased density in tumor and tumor + muscle images compared to images containing muscle. Next, variables were quantified from images of resected primary sarcomas and used to optimize a multivariate model. The sensitivity and specificity for differentiating positive from negative ex vivo resected tumor margins was 82% and 75%. The utility of this approach was further tested by imaging the in vivo tumor cavities from 34 mice after resection of a sarcoma with local recurrence as a bench mark. When applied prospectively to images from the tumor cavity, the sensitivity and specificity for differentiating local recurrence was 78% and 82%. The results indicate that SCA+CT can accurately delineate APFs in heterogeneous tissue, which is essential to enable automated and rapid surveillance of tissue pathology.

Two primary challenges were identified in the work in aim 1. First, while SCA can be used to isolate features, such as APFs, from heterogeneous images, its performance is limited by the contrast between APFs and the background. Second, while it is feasible to create mosaics by scanning a sarcoma tumor bed in a mouse, which is on the order of 3-7 mm in any one dimension, it is not feasible to evaluate an entire human surgical margin. Thus, improvements to the microscopic imaging system were made to (1) improve image contrast through rejecting out-of-focus background fluorescence and to (2) increase the field of view (FOV) while maintaining the sub-cellular resolution needed for delineation of nuclei. To address these challenges, a technique called structured illumination microscopy (SIM) was employed in which the entire FOV is illuminated with a defined spatial pattern rather than scanning a focal spot, such as in confocal microscopy.

Thus, the second aim was to improve image contrast and increase the FOV through employing wide-field, non-contact structured illumination microscopy and optimize the segmentation algorithm for new imaging modality. Both image contrast and FOV were increased through the development of a wide-field fluorescence SIM system. Clear improvement in image contrast was seen in structured illumination images compared to uniform illumination images. Additionally, the FOV is over 13X larger than the fluorescence microendoscope used in aim 1. Initial segmentation results of SIM images revealed that SCA is unable to segment large numbers of APFs in the tumor images. Because the FOV of the SIM system is over 13X larger than the FOV of the fluorescence microendoscope, dense collections of APFs commonly seen in tumor images could no longer be sparsely represented, and the fundamental sparsity assumption associated with SCA was no longer met. Thus, an algorithm called maximally stable extremal regions (MSER) was investigated as an alternative approach for APF segmentation in SIM images. MSER was able to accurately segment large numbers of APFs in SIM images of tumor tissue. In addition to optimizing MSER for SIM image segmentation, an optimal frequency of the illumination pattern used in SIM was carefully selected because the image signal to noise ratio (SNR) is dependent on the grid frequency. A grid frequency of 31.7 mm-1 led to the highest SNR and lowest percent error associated with MSER segmentation.

Once MSER was optimized for SIM image segmentation and the optimal grid frequency was selected, a quantitative model was developed to diagnose mouse sarcoma tumor margins that were imaged ex vivo with SIM. Tumor margins were stained with acridine orange (AO) in aim 2 because AO was found to stain the sarcoma tissue more brightly than acriflavine. Both acriflavine and AO are intravital dyes, which have been shown to stain nuclei, skeletal muscle, and collagenous stroma. A tissue-type classification model was developed to differentiate localized regions (75x75 µm) of tumor from skeletal muscle and adipose tissue based on the MSER segmentation output. Specifically, a logistic regression model was used to classify each localized region. The logistic regression model yielded an output in terms of probability (0-100%) that tumor was located within each 75x75 µm region. The model performance was tested using a receiver operator characteristic (ROC) curve analysis that revealed 77% sensitivity and 81% specificity. For margin classification, the whole margin image was divided into localized regions and this tissue-type classification model was applied. In a subset of 6 margins (3 negative, 3 positive), it was shown that with a tumor probability threshold of 50%, 8% of all regions from negative margins exceeded this threshold, while over 17% of all regions exceeded the threshold in the positive margins. Thus, 8% of regions in negative margins were considered false positives. These false positive regions are likely due to the high density of APFs present in normal tissues, which clearly demonstrates a challenge in implementing this automatic algorithm based on AO staining alone.

Thus, the third aim was to improve the specificity of the diagnostic model through leveraging other sources of contrast. Modifications were made to the SIM system to enable fluorescence imaging at a variety of wavelengths. Specifically, the SIM system was modified to enabling imaging of red fluorescent protein (RFP) expressing sarcomas, which were used to delineate the location of tumor cells within each image. Initial analysis of AO stained panels confirmed that there was room for improvement in tumor detection, particularly in regards to false positive regions that were negative for RFP. One approach for improving the specificity of the diagnostic model was to investigate using a fluorophore that was more specific to staining tumor. Specifically, tetracycline was selected because it appeared to specifically stain freshly excised tumor tissue in a matter of minutes, and was non-toxic and stable in solution. Results indicated that tetracycline staining has promise for increasing the specificity of tumor detection in SIM images of a preclinical sarcoma model and further investigation is warranted.

In conclusion, this work presents the development of a combination of tools that is capable of automated segmentation and quantification of micro-anatomical images of thick tissue. When compared to the fluorescence microendoscope, wide-field multispectral fluorescence SIM imaging provided improved image contrast, a larger FOV with comparable resolution, and the ability to image a variety of fluorophores. MSER was an appropriate and rapid approach to segment dense collections of APFs from wide-field SIM images. Variables that reflect the morphology of the tissue, such as the density, size, and shape of nuclei and nucleoli, can be used to automatically diagnose SIM images. The clinical utility of SIM imaging and MSER segmentation to detect microscopic residual disease has been demonstrated by imaging excised preclinical sarcoma margins. Ultimately, this work demonstrates that fluorescence imaging of tissue micro-anatomy combined with a specialized algorithm for delineation and quantification of features is a means for rapid, non-destructive and automated detection of microscopic disease, which could improve cancer management in a variety of clinical scenarios.

THE GREAT RUSSIAN PIANO TRADITION: SELECTED REPERTOIRE BY TCHAIKOVSKY, RACHMANINOFF AND PROKOFIEV

This dissertation explores representative piano music by three great Russian composers: Tchaikovsky, Rachmaninoff and Prokofiev. The areas of research include: 1) the short character piece; 2) the Russian piano transcription tradition; 3) the concerto and sonata cycle; 4) extra-musical imagery; 5) the influence of popular and dance music of the period. Perhaps the most important result of this research is learning how the art of incorporating a singing quality at the piano stands at the center of Russian pianistic heritage. The first recital features compositions by Sergei Prokofiev. The Seventh Sonata exhibits rebellious, uncompromisingly dissonant treatment of its musical content. Ten Pieces from “Cinderella” shows an ascetic approach to piano texture - a common characteristic in Prokofiev’s late works. The Third Concerto is Prokofiev’s masterpiece in the genre. One of the 20th century’s most performed concerti, it overflows with pianistic challenges. For my second dissertation recital, I have chosen Peter Ilich Tchaikovsky’s The Seasons. These short character pieces were inspired by literary sources. The text portrays Russian rural life, nature, moments of intimate reflection, and imaginary experiences and impressions. Tchaikovsky’s gift as a melodist and remarkable musical individualist is represented in his two Nocturnes as well as in the Nutcracker Suite, masterfully transcribed by Mikhail Pletnev. The final program features Sergei Rachmaninoff’s Ten Preludes, Op. 23, regarded as a culmination of the turn-of-the-century grand Russian pianistic style. The Fantasy Pieces helped establish Rachmaninoff’s reputation as a pianist-composer, a profoundly lyrical poet of the piano. The three Rachmaninoff transcriptions, the Minuet, the Hopak and the Polka de W.R. preserve the spirit of the Golden Era’s musical salon. These pieces were written to delight and dazzle audiences with their bold character, musical taste, virtuosic tricks and technical finesse. The three recitals comprising this dissertation were presented in Gildenhorn Recital at the University of Maryland School of Music on November 13, 2010, April 11, 2011 and February 27, 2012. The recitals were recorded on compact discs and are archived within the Digital Repository at the University of Maryland (DRUM).

Bridging the Divide Between the Soloist and the Ensemble Player

In conservatories and music schools, the general practice for an aspiring pianist is to focus on solo performance learning mainly solo repertoire. With the advent of the advanced degree in collaborative piano, pianists could submerge themselves in the study of duo sonatas, larger chamber music ensembles, and art song. The appearance of this degree was an important step in the development of pianists, as this kind of work requires specific training and focus to master the vast repertoire involved. However it also more clearly brought out the invisible divide separating the solo pianist from the collaborative pianist, a.k.a. the accompanist. While geniuses such as Bach, Beethoven and Brahms were known to compose and perform all types of music, the appearance of super stars such as Liszt and Paganini helped bring into being the term accompanist and since then music world has tacitly embraced this divide. The goal of my dissertational study is to show that this divide need not exist. The three recitals which comprise this dissertational project were all performed at the University of Maryland, the first on 12 November 2010 at Gildenhom Recital Hall, the second at Ulrich Recital Hall on 10 September 2011, and the third at Gildenhorn Recital Hall on 11 November 2011. The repertoire included Rachmaninoff Prelude in g# minor op. 32 no. 12 and Etude-Tableaux in Eb minor op. 29 no. 5, Brahms Sonata for Piano and Violin in d minor op. 108, Mendelssohn Piano Trio in d minor op. 49, Chopin Sonata No.2 in Bb minor, Franck Sonata for Piano and Violin, Prokofiev Piano Concerto no. 2 in g minor op. 16 with pianist Elizabeth Brown as orchestra, Beethoven Sonata for Piano and Violin in A op 47 (Kreutzer), and Paul Schoenfield Cafe Music. All works with violin and cello were performed with violinist Rebecca Racusin, and cellist Devree Lewis. The recitals were recorded on compact discs and are archived within the Digital Repository at the University of Maryland(DRUM).